The present study was carried out in order to set up a standardized quantitative assay for spontaneous micronuclei in bovine lymphocytes. For this purpose the cytokinesis-block micronucleus (MN) method, originally proposed by Fenech and Morley (1985) for human lymphocytes, was applied to peripheral blood lymphocytes of 20 healthy cows of Italian Friesian breed. The results demonstrate that the optimal concentration of cytochalasin B to obtain the highest frequency of binucleated cells (mean = 400.26 +/- 23.76/1000 cells scored) was 6 micrograms/ml. The baseline frequency of spontaneous MN formation in 500 binucleated cells was 12.3 +/- 4.1, i.e., 3 times higher than that reported in human lymphocytes (Fenech and Morley, 1985; Scarfi et al., 1991). The possible reason(s) for this difference (sensitivity to cytochalasin B, chromosome number, environmental genotoxic pollutants) is discussed.